Method and sensor for detecting presence or absence of a contaminant

ABSTRACT

The present invention relates in one aspect to a method of detecting a contaminant in a measurement chamber ( 201 ) of a sample analyzer ( 200 ). The sample analyzer ( 200 ) comprises an optical sensor with a sensor layer ( 205 ) comprising a luminophor ( 201 ), wherein the sensor layer ( 205 ) has a sensor surface ( 206 ) forming an interface to the measurement chamber ( 201 ). The method comprises steps of: filling the measurement chamber with a fluid sample; applying a stimulus to the luminophor in the sensor layer; detecting luminescence emitted from the luminophor in the sensor layer in response to the stimulus as a function of time; obtaining a time sequence of measurement values for the detected luminescence; based on the time sequence, determining an actual value of a first parameter and an actual value of a second parameter, wherein one of the first and second parameters is sensitive to a change in refractive index across the interface between the sensor layer and the measurement chamber, and wherein the other one of the first and second parameters is not sensitive to said change in refractive index across the interface between the sensor layer and the measurement chamber; developing an expected value for the second parameter based on the actual value of the first parameter; comparing the expected value for the second parameter to the actual value of the second parameter; and determining the presence (or absence) of a contaminant based on the comparison. In a further aspect, a sample analyzer configured for detecting contaminants in the measurement chamber using embodiments of the above method is provided.

The present invention relates in one aspect to a method of detecting acontaminant in a measurement chamber of a sample analyzer, wherein thesample analyzer comprises an optical sensor with a sensor layercomprising a luminophor, wherein the sensor layer has a sensor surfaceforming an interface to the measurement chamber.

In a particular aspect, the present invention relates to a method ofdetecting a contaminant in a body fluid analyzer, such as a bloodanalyzer, the analyzer comprising an optical sensor configured formeasuring a body fluid parameter, such as the partial pressure of oxygenin a blood sample, and to a body fluid analyzer, such as a bloodanalyzer, comprising a measurement chamber with such an optical sensor,and a signal processor, the analyzer being configured for detecting acontaminant in the measurement chamber.

According to a further aspect, the invention relates to an opticalsensor for detecting a contaminant in the measurement chamber.

According to a yet further aspect, a computer-implemented method ofdetecting a contaminant in a sample analyzer, and a correspondingsoftware product that can be loaded into a signal processor of a sampleanalyzer are provided. Also in this aspect, the sample analyzer may be abody fluid analyzer, such as a blood analyzer for analyzing e.g. a wholeblood sample.

BACKGROUND OF THE INVENTION

Analyzers for measuring physical parameters of analytes in a liquidsample by means of respective analyte sensors are widely used in variousindustries, such as food industry, environmental industry, as well asmedical and clinical industry. To ensure both accurate and preciseresults, the performance of such analyzers and the associated sensors iscontinuously scrutinized. This typically includes both detailedcalibration and quality control procedures using standardized referenceliquids including the respective analytes in well-defined compositions.The accurate and precise operation of analyzer systems is of particularimportance in clinical analysis applications for analyzing physicalparameters of analytes in bodily fluids, such as whole blood. Inaddition to the accuracy, precision, and reliability requirements, suchanalyzer systems for clinical applications are also subject to furthercritical constraints, such as a short time to obtaining a measurementresult, and the capability of providing the highly reliable results fromvery small sample volumes.

The combination of all these constraints is particularly relevant inblood analyzers. Blood analyzers provide measurements of variousparameters for analyzing the blood of a mammal subject, e.g. forestablishing and/or monitoring a biological condition of the subject.Typically, the mammal subject is a human patient. In a variety ofinstances, it is desirable to measure e.g. the partial pressure of bloodgasses in a whole blood sample of the mammal subject, concentrations ofelectrolytes and metabolites in the blood sample, as well as thehematocrit value of the blood sample. For example, measuring pCO₂, pO₂,pH, Na⁺, K⁺, Ca²⁺, Cl⁻, glucose, lactate, creatinine, urea andhemoglobin and hemoglobin-derivate values are primary clinicalindications in assessing the condition of a medical patient. A number ofdifferent analyzers currently exist for making such measurements. Suchanalyzers are able to perform precise measurements in order to providethe most meaningful diagnostic information.

In order to use as little of the patient's blood as possible in eachanalysis performed, the measuring chamber, which is employed to analyzea blood sample, is preferably relatively small. Performing bloodanalysis using a small blood sample is important when a relatively largenumber of samples must be taken in a relatively short amount of time orif the volume of blood is limited, as in neonates. For example, patientsin intensive care require a sampling frequency of 15-20 per day forblood gas and clinical chemistry measurements, leading to a potentiallylarge loss of blood during patient assessment. Furthermore, in order tolimit the number of tests which must be performed it is desirable togather as much information as possible upon completion of each test.Furthermore, for the same reasons, it is important that the measurementsand corresponding analysis results obtained from these measurements arereliable. Each measurement is therefore typically subject to acalibration and/or quality control procedure using different rinsing,calibration and/or reference liquids and the measurement chamber isthoroughly rinsed after each measurement to avoid contamination of anysubsequent measurements.

However, a common issue in blood analyzers, in particular in systemswith very small measurement chambers, is due to the presence of clots inwhole blood samples. The clots may result in the formation of plugsimpeding, obstructing or even completely blocking the fluid passages ofthe measurement chamber. Such clots may severely affect the measurementsor even cause damage to the measurement chamber/sensor assembly. Knownsystems may therefore monitor the filling and discharge procedures forabnormalities in order to e.g. generate an alarm, stop the fluidhandling infrastructure from feeding further fluid to the measurementchamber, and requesting a rinse and/or initiating an automated rinsingprocedure. For example, the filling of the measurement chamber may bemonitored by liquid sensors for detecting the passage of a liquidinterface at an inlet upstream of the measurement and the correspondingsubsequent occurrence of the liquid at an outlet downstream of themeasurement chamber after an expected filling time. Unexpected behavior,e.g. expiry of the expected filling time without positive detection ofthe liquid interface at the outlet liquid sensor, may result in an alarmand/or initiation of a rinsing/maintenance procedure. Furthermore, bydesigning a simple flow path through the measurement chamber theformation of deposits can be counteracted and rinsing/wash-out can befacilitated.

While such strategies for detecting the presence of a clot in themeasurement chamber are implemented and proof to be most helpful for areliable operation of blood analyzer systems, it has been observed bythe inventors that not all artifacts may be accounted for by thesestrategies and detection techniques. The inventors have indeedidentified that further artifacts may arise due to clots, which arenormally not detectable by the known clot detection routines that arebased on e.g. flow behavior. Clots that do not noticeably affect thefilling and discharge flow in the measurement chamber may neverthelesscause a severe distortion of the physical parameters of at least some ofthe analytes for a given sample, thus leading to erroneous analysisresults. Therefore, there is a need for rapidly and reliably detectingany such additional causes of potential artifacts in blood analyzers inorder to ensure accuracy and precision of the measurements and avoid thewaste of valuable patient blood. Furthermore, such additional artifactsmay also occur more generally in liquid sample analyzers. To addresssuch additional causes of artifacts, the inventors have, in co-pendingpatent applications WO2017/108646A1 and WO2017/108647A1, which arehereby incorporated by reference, suggested different techniques fordetecting clots by analyzing deviations of sensor response from anexpected behavior. The clot detection disclosed in WO2017/108646A1relates to a method based on an expected change i.e. a linear regressionof the measurement results from at least two sensors, where deviationfrom the expected change is indicative of a clot. Clot detection inWO2017/108647A1 discloses a method in which a clot may be seen as areservoir with a capacity for the uptake and emission of analyte,thereby causing pollution by acting as an analyte source or as ananalyte sink whenever there is a gradient in the analyte concentrationbetween the clot and the surrounding liquid sample.

However, further techniques for the detection of fouling or othercontaminants in a measurement chamber of a liquid sample analyzer arestill desired in order to further improve the reliability of theobtained measurement results, in particular when using very small samplevolumes and/or when using samples with limited availability as alreadydiscussed above.

One type of analyte sensors are optical sensors with a sensor layer incontact with the measurement chamber. The sensor layer is sensitive toan amount of an analyte present in the fluid sample that is provided inthe measurement chamber. The optical sensor further comprisesinstrumentation for the optical readout of the sensor layers' responseto the presence of the analyte. The readout means typically comprisemeans for providing a stimulus to the sensor layer, optical elements,such as lenses and/or optical wave guiding components for collectingradiation emitted from the sensor layer in response to the stimulus, andfurther for transferring the collected luminescence radiation todetection means of the optical sensor. The stimulus is typically aradiation source, such as a laser or light emitting diode (LED),arranged and configured to provide optical probing radiation to thesensor layer. Optical sensors may, for example be of the fluorescencequenching type. The sensor layer of a luminescence quenching type sensorcomprises a luminophor that is excited by the stimulus provided, such asexcitation radiation guided to the sensor layer. The excited luminophorrelaxes, amongst others, along radiative pathways, thereby emittingluminescence, which upon termination of the stimulus decays with acharacteristic lifetime. The luminophor is selected such that theluminescence is quenched by the presence of the analyte acting as aso-called quenching agent. As a consequence, the characteristic lifetimeof the luminescence emitted by the sensor layer depends on the amount ofthe analyte present in the sensor layer. An increase in concentration ofthe luminescence quenching analyte results in a decrease of the observedluminescence lifetime, whereas a decrease in concentration results in anincreased lifetime. In order to provide proper results, the measurementis typically conducted at equilibrium conditions, i.e. the fluid sampleis provided in the measurement chamber in such a manner that theconcentration of the analyte in the sensor layer corresponds to theconcentration in the sample, e.g. through diffusive exchange of analytebetween the sensor layer and any fluids presented in the measurementchamber. The optical sensor may further comprise optional means foroptically selecting and/or analyzing the radiation collected from thesensor layer, such as optical filters and/or optical amplifiers, beforethe light is received by the detection means. The detection meansconvert the detected luminescence radiation to a corresponding signal.The optical sensor is thus, configured to provide a signalrepresentative of the amount of the analyte for which it is sensitized.The signal from the optical sensor is then typically provided toprocessing means in the sample analyzer for analog and/or digital signalprocessing, passed to further storage means for storage as measurementdata, displayed and/or presented as analysis results at an output.

To give an example of a sensor, which, when used in a blood analyzer, isof particular importance for patient health and safety is the sensordetermining the partial pressure of oxygen (pO₂). Such optical detectorsfor blood gas measurements, such as for pO₂-measurements, are forexample known from U.S. Pat. No. 5,564,419A. The results obtained frompO₂-measurements can directly affect the treatment of thepatient—particularly in emergency or intensive care situations. The pO₂sensor may be an optical sensor with a sensor layer comprising aluminophor sensitive to the presence of oxygen in the sample. Acontamination of the pO₂ sensor of a blood analyzer will affect themeasurement and thus increase the risk for wrong treatment of thepatient.

Accordingly, there is a further need for rapidly and reliably detectingany such additional causes of potential artifacts in sample analyzers inorder to ensure accuracy and precision of the measurements. Inparticular, a rapid, if not immediate feedback on the reliability ofmeasurements in sample analyzers, such as for measuring body fluidparameters, is desired in order to validate these measurements withoutdelay.

Object of the present invention is therefore to provide a further methodof detecting contamination in a measurement chamber with improvedsensitivity and/or response time, and a system adapted to perform suchdetection method with improved sensitivity and/or response time.According to a further aspect, a further object is to provide adetection scheme allowing for an improved performance for the rapid andreliable detection of invalidating artifacts stemming from measurementchamber contamination.

SUMMARY OF THE INVENTION

A first aspect of the invention relates to a method of detecting acontaminant in a measurement chamber of a sample analyzer, wherein thesample analyzer comprises an optical sensor with a sensor layercomprising a luminophor, wherein the sensor layer has a sensor surfaceforming an interface to a fluid sample in the measurement chamber, themethod comprising the steps of:

-   -   filling the measurement chamber with a fluid sample;    -   applying a stimulus to the luminophor in the sensor layer;    -   detecting luminescence emitted from the luminophor in the sensor        layer in response to the stimulus as a function of time;    -   obtaining a time sequence of measurement values for the detected        luminescence;    -   based on the time sequence, determining an actual value of a        first parameter and an actual value of a second parameter,        wherein one of the first and second parameters is sensitive to a        change in refractive index across the interface between the        sensor layer and the measurement chamber, and wherein the other        one of the first and second parameters is not sensitive to said        change in refractive index across the interface between the        sensor layer and the measurement chamber;    -   developing an expected value for the second parameter based on        the actual value of the first parameter;    -   comparing the expected value for the second parameter to the        actual value of the second parameter; and    -   determining the presence of a contaminant or determining the        absence of a contaminant, based on the comparison.

The sample space of the measurement chamber is filled so as to contactthe sensor surface with the sample fluid, and with the purpose ofperforming a measurement on the fluid sample. It is an important meritof the present invention to recognize and address the significance ofsmall contaminant amounts deposited on the surface of the sensor layerforming the interface of an optical sensor to the measurement chamber.The present method allows for detecting the presence of contaminants inthe sample space of the measurement chamber. More particularly, themethod is sensitive to the presence of contaminants on the sensorsurface of the optical sensor and capable of providing immediatefeedback on the contamination state of an optical sensor, and theadjacent sample space. Moreover, the contaminant detection is suited forquality monitoring purposes by continuously analyzing the time-resolvedluminescence data and providing almost instantaneous feedback to themeasurement system/user on the quality of the measurement and on theinternal state of the sample analyzer used for analyzing the fluidsample.

The term ‘contaminant’ as used herein refers to any substance in a fluidsample that potentially may interfere with a measurement to be performedon the fluid sample. The term ‘fluid’ as used herein refers to both‘liquid’ and ‘gas’. Examples of contaminants may be droplet remaindersof a previously measured liquid sample, precipitations on themeasurement chamber walls, and/or bubbles occurring in a liquid samplee.g. due to an improper filling of the measurement chamber with a liquidsample, or the like. The present method is particularly useful in thecontext of a re-usable measurement chamber, i.e. a multi-use measurementchamber, which after single use, i.e. after performing a measurementcycle on a single fluid sample, is not discarded, but emptied, rinsed,and filled again with a new fluid sample. However, it is alsoconceivable to use the contamination detection method in the context ofa single-use measurement chamber, which after a measurement cycle on afluid sample is discarded, e.g. for ensuring cleanliness and/or properfilling of the single-use measurement chamber, before the actualmeasurement is performed.

The sensor layer is transparent and has a sensor surface forming a frontside interface towards the sample space. The front side interface is incontact with the fluid sample. Optical probing is performed from theback side, i.e. from the side facing away from the sample space. Astimulus is given, typically in the form of excitation light that isdirected to the sensor layer from the backside, so as to produce anexcited fraction of the luminophor in the sensor layer. The excitedluminophor molecules relax back to a ground state under the emission ofluminescence light, which may also be observed from the back side. Theoptical sensor therefore further comprises instrumentation for detectingand registering luminescence emitted from the luminophor, and thus toobserve the sensor layer response to the applied stimulus.

The parameter, which does not dependent on the change in refractiveindex across the interface between the sensor layer and the measurementchamber, may be referred to as an “intrinsic” parameter. An “intrinsic”parameter is thus representative of the “intrinsic”characteristics/properties of the relaxation process of the luminophordye, which involves the radiative recombination at the origin of thedetected luminescence radiation—essentially independent of the opticalstructure in which the luminophor is embedded, in particular invariantwith respect to changes in the optical properties of the interface withthe measurement chamber at the sensor surface.

The luminescence radiation is generated within the sensor layer.Radiation emitted in the direction towards the sample space is reflectedat the optical interface between the transparent sensor layer and thesample space. The back reflected luminescence radiation thereforecontributes to the radiation that is collected for detection. Thereflection depends on the optical properties of the interface, which onone side is determined by the optical properties of the sensor layer andon the other side is determined by the optical properties of thesubstances in the sample space, in contact with the sensor surface. Theback-reflected radiation therefore comprises information about theoptical properties of any substances in contact with the sensor surface.The substances may thus be distinguished by their optical properties.More particularly, any contaminants and the fluid sample are thusdistinguishable by any differences in refractive index.

The parameter, which does, at least partly, dependent on the change inrefractive index across the interface between the sensor layer and themeasurement chamber, may be referred to as an, at least partly,“extrinsic” parameter. The “extrinsic” parameter is thus sensitive tothe part of the detected luminescence radiation that is sensitive to theoptical characteristics of the structure in which the luminophor isembedded and from which it is collected. In particular, the extrinsicparameter is sensitive to changes in the optical properties of theinterface between the sensor layer and the measurement chamber.

For determining the presence or absence of a contaminant on the sensorsurface, actual values for the first and second parameters are obtainedfrom a time sequence of optical probing measurements using the opticalsensor. The first and second parameters are thus physically relatedthrough the luminescence from which they originate. One of the two mayhowever, vary according to the contamination state, which influences theoptical properties of the interface between the sensor layer and thesample space as described above. To facilitate distinction betweendifferent contamination states of the optical sensor surface, referenceinformation is provided. Reference measurements on one or more referencefluids are prepared beforehand to establish a unique relation betweenthe first and second parameter for each of the one or more referencefluids. Suitable reference fluids may be any liquid and/or gas sampleswith known optical characteristics, such as an aqueous solution with aknown composition as commonly used in medical sample analyzers for QC orcalibration purposes, air prepared to a predefined state, nitrogen, anoble gas, such as argon, or the like. The reference measurements areperformed for one or more known contamination states. Typically, thereference measurements are performed with a clean sensor surface, i.e.in the absence of any contamination on the sensor surface, the surfacethus only being in contact with the respective reference fluid. Thereference measurements provide reference information that may be storedor in any other way made accessible to a processor for later use. Thereference information may be consolidated and/or stored in any suitableform, for example tabulated, in a parametrized relation, and/or as oneor more coefficients in an equation describing the relation between thefirst and second parameters for at least one known contamination state,which most preferably is the above-described clean state of the sensorin respect of at least one reference fluid. The sample analyzer is thusconfigured for developing an expected value for one of the first andsecond parameters based on actual values for the other one of the firstand second parameters. The developed expected value is then compared tothe actual value of the other one of the first and second parameters. Incase the two values match as expected, the absence of contamination isdetermined. In case a mismatch between the expected value and the actualvalue of the other one of the first and second parameters is observed, acontamination of the sensor surface can be concluded.

Advantageously, according to some embodiments, the first parameter isthe one that is not-dependent on the optical properties of the interfaceat the sensor surface (“intrinsic”), and the second parameter is the onethat is dependent on said interface (at least partly “extrinsic”).Further advantageously, the expected value is determined based on theactual value of the intrinsic parameter. Thereby, it is achieved thatthe expected value of the second parameter calculated on the basis ofthe first, intrinsic parameter is less prone to artefacts stemming fromthe optical structure of the detection instrumentation.

Any suitable luminophor may be used, in combination with an opticalprobing technique that allows for determining a first parameter and asecond parameter of the luminescence radiation emitted in response to astimulus applied to the luminophor, wherein one of the first and secondparameters depends on the optical properties of the sensor layer/sampleinterface, and the other one does not as required above.

The optical sensor is typically arranged for measuring a particularphysical parameter for an analyte in a fluid sample present in themeasurement chamber. For example, the optical sensor may be arranged ina sensor cassette comprising a measurement chamber with an inlet and anoutlet, and multiple analyte sensors, said analyte sensors each beingadapted to measure a respective parameter in respect of an analyte. Theoptical sensor used for the detection of contaminants may be one of theanalyte sensors. However, it is also conceivable that the optical sensoris dedicated to the detection of contaminants only. Advantageously, theoptical sensor is placed in the measurement chamber in such a way thatthe probability of detecting a contaminant is enhanced, e.g. at alocation where contaminants are more likely to be detected, or at alocation where contaminants tend to accumulate. Further advantageously,the optical sensor used for the detection of contaminants may be placedat the inlet. This has the advantage that any contaminant introducedinto measurement chamber from the outside has to pass by the opticalsensor. Furthermore, flow conditions around an inlet port may entail anenhanced probability of contaminants depositing on the measurementchamber sidewalls. By placing the optical sensor used for the detectionof contaminants, the probability of capturing a contaminant may thus beenhanced as compared to other locations. Alternatively or in additionthereto, according to further considerations, an optical sensor to beused for the detection of contaminants may also be placedopposite/vis-à-vis/of another sensor, e.g. a sensor that for some reasoncannot itself be used for the detection of contaminants, that isparticularly sensitive to the presence of contaminants, and/or that hasa tendency to attract contaminants. For example, an optical sensor forthe detection of contaminants may be placed at a location in themeasurement chamber that is opposite to a sensor for measuring pCO₂.According to yet further considerations, an optical sensor for thedetection of contaminants, wherein the optical sensor is also an analytesensor for measuring a parameter in respect of a specific analyte, mayalso be placed so as to optimize for the analyte measurement. In casethere is a conflict between these considerations, the skilled person candetermine a location as a compromise, or according to furtherpreferences/priorities. Depending on the sensor type, optical probingmay involve a luminophor in the transparent sensor layer that issensitive to the analyte, for example a luminophor that may be probedfor the presence of the analyte using known luminescence quenchingtechniques. Examples for luminescence quenching dyes are generally knownin the art. For example, porphyrin compounds, such as aryl-substitutedtetrabenzoporphyrin, palladium porphyrin (e.g. PdTPP, or PdTFPP) fordetecting the presence of oxygen acting as a quenching agent for theseporphyrin-compound based dyes. The present method for detectingcontaminants has the advantage that it may probe the radiation responsefrom the same luminophor that is also useful for probing for theanalyte. The present method is therefore, in a synergistic way,particularly useful for detecting contaminants on the surface of asensor that is adapted for measuring analytes by luminescence-probingtechniques and contaminant detection may thus be easily implemented insuch a luminescence probing set-up.

The method for detecting contaminants may be used at any time before,immediately before, during, and/or after a measurement is performed, inorder to provide instant feedback on the contamination state of thesensor surface. In certain cases, the contamination state of the opticalsensor may also reflect the general contamination state of themeasurement chamber. Thereby, instant information on the quality andreliability of a measurement is available. The instant availability ofthe contamination state information also allows for immediately takingcorrective measures, such as to clean the sensor surface, themeasurement chamber as a whole, before a new measurement is performed,or in some cases to apply adequate corrections at a data analysis level.Acting immediately upon the occurrence of a contamination state avoidswaste of valuable sample material, that otherwise would result from abelated, retroactive invalidation of the sample results. Immediatecorrective action is also important for providing the correct care hereand now, e.g. in emergency or intensive care, where the difference canbe live saving.

The present invention is particularly useful for the detection ofcontaminants, such as clots or bubbles, in the measurement chamber of amedical sample analyzer, and further for verification of the presence ofa suspected contamination or after conclusion of measures for theremoval of a previously detected contamination. The detection result maybe used as a part of self-control routines of the medical sampleanalyzer, or may be requested by a user or otherwise be triggeredexternally. The detection result may further trigger an alarm or errorstate of the medical sample analyzer, and may also be used to invoke acontaminant removal procedure and/or request external service,maintenance or replacement of a faulty measurement chamber if removal ofthe contamination proofs unsuccessful.

Further according to some embodiments of the method, the presence of acontaminant is determined if the difference between the actual value ofthe second parameter and the expected value for the second parameter isabove a threshold, and/or wherein absence of a contaminant is determinedif the difference between the actual value of the second parameter andthe expected value for the second parameter is below the threshold.Thereby a more reliable distinction between the presence and absence ofcontaminants is achieved. Furthermore, a threshold can be set so as todiscriminate between insignificant and significant contaminationaccording to the significance of an observed mismatch between thedeveloped expected value and the corresponding actual value for thevalidity of measurements to be performed using the optical sensor.

Further according to some embodiments of the method, the fluid sample isan aqueous liquid. When the fluid sample is an aqueous solution, themethod is particularly useful for the detection of gas phasecontaminants, such as bubbles. For example, the sample may be an aqueoussolution or other water-based liquid, such as for medical parameteranalysis, blood, urine, or related calibration/QC solutions, and thecontaminant may be a bubble of gas adhering to the sensor surface. Thelarge difference in refractive index between the gas bubble and theaqueous sample allows for a reliable detection of the contaminantindicating a deviant result using the method on an aqueous sampleliquid. Advantageously, the method of detecting a bubble is performedduring or in connection with the preparation/presentation of a liquidsample in the measurement chamber, wherein a bubble having a refractiveindex comparable to a gas sample can be detected by filling themeasurement chamber with the liquid sample, and performing the opticalbubble detection method as described herein. Thereby an indication for aproper filling of the measurement chamber can be verified. This isparticularly an advantage, when the volume of the measurement chamber isin very small, such as in an elongate channel shaped chamber withdimensions in directions transverse with respect to a principal axis ofthe chamber in the mm and sub-millimeter range.

Further according to some embodiments of the method, the fluid sample isa liquid with a refractive index between 1.20 and 1.50, such as between1.25 and 1.45, between 1.30 and 1.40, or about 1.20; 1.25; 1.30; 1.35;1.40; 1,45; or 1.50. Advantageously, the fluid sample is a liquid with arefractive index between 1.20 and 1.30, between 1.25 and 1.35, between1.30 and 1.40, between 1.35 and 1.45; or between 1.40 and 1.50.Refractive index values are for a given sample analyzer configuration inrespect of the wave lengths of the luminescence radiation emitted by theluminophor in response to the stimulus, and are for temperature rangesat which measurements are typically performed. For example for bodyfluids, the temperature at which the measurements are to be performed istypically specified to a range corresponding to body temperature, suchas between 35° C. and 39° C.; between 36° C. and 38° C.; between 35° C.and 38° C.; between 36° C. and 39° C.; or about 35° C.; 36° C.; 37° C.;38° C. or 39° C.

When using the herein disclosed method on sample liquids with arefractive index within the above-specified ranges a clear distinctionof the sample from a gaseous contaminant at the sensor surface, such asa gaseous contaminant with a refractive index well below 1.10; below1.05; below 1.01; or about 1.00 is achieved.

Further according to some embodiments of the method, the fluid sample isa gas. Another type of contaminant than bubbles in a liquid may bedeposits on the surface of the sensor surface, such as a remainder froma liquid sample from a previous measurement cycle, which maycollectively be referred to as “clots”. When the fluid sample is a gas,the method is particularly useful for the detection of such clots. Clotsmay include droplets, precipitations, and/or may have a gel-likeconsistence with a refractive index corresponding to that of liquids,such as aqueous liquids. Refractive index values in the above-mentionedranges, such as above 1.10, above 1.20, between 1.20 and 1.50, or any ofthe ranges or values mentioned above with respect to liquid samples. Dueto the difference in refractive index between such a clot and a gassample, these clots can be reliably detected in a manner analogue todetecting bubbles of gas in a liquid sample.

Advantageously, the method of detecting a clot is performed during or inconnection with a purging cycle. The purging or rinsing cycle istypically performed after a measurement has been concluded with thepurpose to clean the measuring chamber and prepare the measurementchamber for a new measurement. A clot having a refractive indexcomparable to a liquid sample can be detected by filling the measurementchamber with a reference gas sample, such as an argon or nitrogen gassample, and performing the optical clot detection method as describedherein, prior to filling the measurement chamber with a liquid sample,such as a valuable patient sample. Thereby an additional check of themeasurement chamber is achieved, which allows for instant feedback, andtake immediate corrective action so as to avoid waste of valuablepatient samples.

Further according to some embodiments of the method, the gas has arefractive index of below 1.10; below 1.05; below 1.01, or about 1.

Further according to some embodiments of the method, the refractiveindex of the sensor layer is at least 1.40; between 1.40 and 1.45; atleast 1.45; between 1.45 and 1.50; at least 1.50; between 1.50 and 1.55;or at least 1.55. As mentioned above, the successful detection of acontaminant relies on distinguishing the contaminant and the fluidsample by their refractive index. Preferably, the refractive index ofthe sensor layer differs at least from the refractive index of the fluidsample. Most preferably, the refractive index of the sensor layerdiffers from the refractive indices for both the fluid sample and thecontaminant. Thereby, a reliable detection of the contaminant isachieved.

A typical optical sensor for use in a medical sample analyzer may have asensor layer with a matrix made of a polymer material, such as e.g.cellulose acetate, polyurethane, polycarbonate/silicone copolymer, orpolyvinylchloride (PVC), in which the luminophor is embedded. Matrixmaterials for optical sensors are discussed in detail in the backgroundsection of the international patent application publicationWO2001/004631, wherein further useful matrix materials are disclosed inthe claims of WO2001/004631. WO2001/004631 is hereby enclosed byreference in its entirety. The host material essentially determines therefractive index, which in the case of polymer materials, such as PVC istypically about 1.50, such as in the range from 1.45 and up to including1.55. Unless specified otherwise, refractive index values are for therelevant detection spectral range with respect to the luminescenceradiation emitted by the luminophor in the visible part of the spectrum.In so far a temperature dependence of the refractive index has to betaken into account, the refractive index values refer to temperaturescorresponding to typical temperatures for performing measurements asalready discussed above.

Further according to some embodiments of the method, the step ofapplying a stimulus to the luminophor includes illuminating the sensorlayer with light in an excitation spectral range adapted for excitingthe luminophor. The stimulus is typically applied in a pulsed ormodulated manner with stimulation periods where the stimulus is in anON-state, separated by idle periods where the stimulus is in anOFF-state. This allows for observing time dependent luminescenceresponse of the sensor layer to the stimulus, and more particularly, thepulse and/or step response, of the sensor layer to the stimulus. Typicallight sources used for such optical stimulation may include pulse lasersources or modulated light sources using light emitting diodes (LED).Using laser pulses, very short excitation pulses may be achieved, whichare typically short as compared to lifetime of the luminescenceradiation from the luminophor. LED-based modulated light allow for aless complex, and less costly setup and have longer pulse duration.

Further according to some embodiments of the method, the step ofobtaining a time sequence of measurement values includes measuring theluminescence intensity at a plurality of at least three points in time.A minimum of three points in time is required in order to be able toobtain sufficient independent measurements for determining the actualvalues for the first and second parameters independent of each other.For reasons of noise reduction, a larger number of measurements mayadvantageously be used for the time sequence in order to improvereliability of the obtained values. Good results are for exampleachieved by using between 5-200 points; 5-150 points; 5-100 points;10-200 points; 10-150 points; 10-100 points; 20-200 points; 20-150points; 20-100 points; 30-200 points; 30-150 points; 30-100 points; orat least 5 points, at least 10 points, at least 20 points, at least 30points, at least 40 points, at least 50 points, or at least 60 points intime. An upper limit of the number of measurement points to be acquiredat different times may be subject to an upper limit for the total timeto be spent for the detection procedure. A useful upper limit may be upto 100 points, up to 150 points, or up to 200 points in time.

Further according to some embodiments of the method, the time sequenceof measurement values is obtained for a time window after termination ofthe stimulus. For example, the time window may start immediately upontermination of the stimulus. According to some embodiments, the step ofobtaining the time sequence of measurement values may be triggered bythe termination of the stimulus. By obtaining the time sequence ofmeasurement values after termination of the stimulus, a more simplesubsequent analysis of the time-sequence is achieved. After terminationof the stimulus, the intensity of the luminescence radiation decays. Bystarting the measurement immediately after termination the largestsignal intensity is achieved, thereby reducing noise issues.

Further according to some embodiments of the method, the time sequenceof measurement values is obtained for a time window during applicationof the stimulus. In principle, it is also conceivable to derive usefulactual values for the first and second parameters as required by theinvention from a time sequence during or including a time period whenthe stimulus is applied, as long as any effects of the presence of thestimulus on the luminescence by the detector means and the timedependence thereof, e.g. due to continued excitation and/orre-excitation of luminophor dye molecules by the incident stimulus, istaken into account in the subsequent analysis, for example whendeveloping an expected value for the second parameter from the firstparameter. Furthermore, any additional background radiation or any otherartefacts that may result from the presence of the time-dependentstimulus that is scattered into the detection portion of the opticalsensor will have to be accounted for. The effect of any such artefactsmay, for example, be determined beforehand by measuring thecharacteristics of the sensor response in reference measurements, in theabsence of any contamination. This embodiment is useful in combinationwith the use of a stimulus with a longer duration, such as when using anLED based illumination source.

Further according to some embodiments of the method, the first parameteris the lifetime τ of the luminescence, or a corresponding parameter.According to this embodiment, the first parameter is an “intrinsic”parameter, i.e. a parameter intrinsic to the molecular processesgoverning the relaxation of the excited state of the luminophor, such asthe lifetime τ (tau) of the luminescence decay. The intrinsic parameterthus only depends on the intrinsic interactions occurring in the sensorlayer and does not depend on extrinsic interactions, such as the opticsof the interface between the sensor layer and the fluid sample.Determining the fluorescence lifetime τ from the intensity decay in atime-sequence of luminescence measurements is known and can be directlyimplemented in a signal processor of the optical detector. Thefluorescence lifetime is therefore a simple and reliable implementationfor the first parameter as an intrinsic parameter.

Further according to some embodiments of the method, the secondparameter is the intensity of the luminescence at a given point in time,or a corresponding parameter. According to this embodiment, the secondparameter is the luminescence radiation intensity emitted from thesensor layer and collected by the optical detector at a given point intime, e.g. after termination of the stimulus. The collected intensity isan at least partly “extrinsic” parameter, i.e. a parameter that isinfluenced by factors extrinsic to the recombination processes governingthe relaxation of the excited state of the luminophor. Extrinsic factorsmay include the optical environment of the sensor layer, e.g. the opticsof the interface between the sensor layer and the fluid sample. Keepingall other configuration parameters of the optical detection set-up thesame, the extrinsic parameter, here the luminescence intensity that iscollected by the detector, is therefore sensitive to changes inrefractive index at the sensor surface.

Further according to some embodiments of the method, the luminophor inthe sensor layer is a phosphor with a luminescence lifetime from 1 μsand up to including 1 s; and/or wherein the luminophor in the sensorlayer is a phosphor with a luminescence lifetime of at least 10 μs, atleast 20 μs, at least 30 μs; at least 40 μs; at least 50 μs; at least 60μs; at least 70 μs; at least 80 μs; at least 90 μs; at least 100 μs; atleast 150 μs; or at least 200 μs; and/or wherein the luminophor in thesensor layer is a phosphor with a luminescence lifetime up to andincluding 1 s; 100 ms; 10 ms; 1 ms; 500 μs; 300 μs; 150 μs; 30 μs; or 15μs.

Advantageously according to some embodiments, the luminophor in thesensor layer is a phosphor with a luminescence lifetime from 10 μs andup to including 10 ms, from 10 μs and up to including 1 ms, from 10 μsand up to including 100 μs, from 20 μs and up to including 50 μs, from20 μs and up to including 30 μs.

Advantageously according to some embodiments, the luminophor in thesensor layer is a phosphor with a luminescence lifetime of at least 10μs, at least 20 μs, at least 30 μs; at least 40 μs; at least 50 μs; atleast 60 μs; at least 70 μs; at least 80 μs; at least 90 μs; at least100 μs; at least 150 μs; or at least 200 μs; and/or wherein theluminophor in the sensor layer is a phosphor with a luminescencelifetime up to and including 1 s; 100 ms; 10 ms; 1 ms; 500 μs; 300 μs;150 μs; 30 μs; or 15 μs.

For the purpose of these embodiments, the lifetime of the luminophoremission is considered in the absence of luminescence quenching effects.For a luminophor adapted for luminescence quenching measurements inrespect of a quenching agent (the specific analyte), the lifetime isconsidered in the absence of the quenching agent. Furthermore, theluminophor emission lifetime is considered under relevant measurementconditions in the context of the application of the contaminationdetection method, as easily determined by a skilled person. For example,in the case of medical analysis equipment, such as point-of-care orlaboratory equipment, the relevant temperature is typically specified toa range corresponding to body temperature, such as between 35° C. and39° C.; between 36° C. and 38° C.; between 35° C. and 38° C.; between36° C. and 39° C.; or about 35° C.; 36° C.; 37° C.; 38° C. or 39° C.

Further according to some embodiments of the method, the step of fillingthe measurement chamber includes bringing the fluid sample in adiffusive equilibrium with the sensor layer, at least with respect toone analyte. Bringing the fluid sample into a diffusive equilibriumstate may require a time delay as a part of the filling step. Thereby,artefacts arising from a non-equilibrium concentration distributionbetween the sample and the sensor layer, where the analyte is measured,may be avoided. Thereby, a simplified and reliable contaminant detectionprocedure is achieved.

Advantageously according to some embodiments of the method, the opticalsensor is adapted to measure one or more analytes in a fluid sample soas to determine a corresponding parameter of the analyte, such as pH,concentrations of electrolytes, concentrations of metabolic factors orconcentrations of enzymes. The fluid sample may be a biological sample,such as a body fluid, i.e. a physiological fluid.

Examples of biological samples may include both liquid and gas samples.Liquid samples may be selected from the group of blood, diluted orundiluted whole blood, serum, plasma, saliva, urine, cerebrospinalliquid, pleura, synovial liquid, ascites liquid, peritoneal liquid,amniotic liquid, milk, dialysis liquid samples, or the like, as well asany quality control materials and calibration solutions used in analyzerequipment for measuring any of these fluids. Gaseous samples may includerespirator gas, expiratory air, or the like, as well as any qualitycontrol and calibration materials used in analyzer equipment formeasuring any of these fluids. The sample may be treated prior totesting in order to make it more amenable to being tested. Pretreatmentmethods may include dilution, filtration, concentration, extraction,removal or inactivation of components which might interfere with theresults, and addition of reagents. Examples of other biological samplesinclude fermentation broths or microbial cultures, waste water, foodproducts, and the like.

Examples of parameters in respect of analytes, which may be determinedby means of the optical sensor of the invention include: pO₂, pCO₂, pH;concentrations of electrolytes such as Li⁺, Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl⁻,HCO³⁻ or NH₃ (NH₄ ⁺); concentrations of metabolic factors, such asglucose, creatinine, urea (BUN), uric acid, lactic acid, pyruvic acid,ascorbic acid, phosphate or protein; concentrations of enzymes such aslactic acid dehydrogenase, lipase, amylase, choline, esterase, alkalinephosphatase, acid phosphatase, alanine amino transferase, aspartate,amino transferase, or creatinine kinase.

Further according to some embodiments of the method, the optical sensoris adapted for measuring a partial pressure of a gas fraction in thefluid sample, such as pO₂, or pCO₂.

Further according to some embodiments of the method, the sensor layer isadapted for the diffusive uptake of an analyte from the fluid sample,and the luminophor in the sensor layer is susceptible to luminescencequenching, due to the presence of the analyte in the sensor layer.

Thereby, the analyte measurement and the contaminant detection can becombined in a synergistic manner. Using luminescence quenchingdetection, a sensitive and precise method of determining concentrationsof specific analytes acting as quenching agent for the luminophor in thesensor layer is provided. Advantageously, the luminescence quenchingmeasurements may be evaluated using a Stern-Volmer-type analysis in aknown manner. At the same time, using the present invention, theluminophor emission can be used for reliably detecting contamination onthe sensor surface. The contamination detection can be achievedessentially instantly, thus allowing corrective measures to be takenimmediately, such as compensating for artefacts resulting from any suchcontamination and/or attempting to remove the contaminant. As aconsequence an enhanced precision of the optical sensor measurement isachieved.

Further according to some embodiments of the method, the optical sensoris adapted to determine a parameter of the fluid sample in respect ofone or more analytes.

Further according to some embodiments of the method, the parameter ofthe fluid sample in respect of one or more analytes is selected from thegroup of: pO₂, pCO₂, pH; concentrations of electrolytes such as Li⁺,Na⁺, K⁺, Ca²⁺, Mg²⁺, Cl⁻, HCO³⁻ or NH₃ (NH₄ ⁺); concentrations ofmetabolic factors, such as glucose, creatinine, urea (BUN), uric acid,lactic acid, pyruvic acid, ascorbic acid, phosphate or protein;concentrations of enzymes such as lactic acid dehydrogenase, lipase,amylase, choline, esterase, alkaline phosphatase, acid phosphatase,alanine amino transferase, aspartate, amino transferase, or creatininekinase.

Further according to some embodiments of the method, the fluid sample isa liquid, such as a body liquid, i.e. a physiological liquid.Correspondingly, a sample analyzer for use in performing the method isadvantageously adapted to analyzing parameters of liquid samples, suchas body liquids, i.e. physiological liquids. A medical sample analyzermay thus advantageously include a liquid handling system comprisingvalves, conduits, and/or pumping/transfer means, for controlling liquidflow, such as for filling and emptying of the measurement chamber withthe liquid sample—preferably in an automated manner.

Further according to some embodiments of the method, the fluid sample isa liquid sample selected from the group of blood, diluted or undilutedwhole blood, serum, plasma, saliva, urine, cerebrospinal liquid, pleura,synovial liquid, ascites liquid, peritoneal liquid, amniotic liquid,milk, dialysis liquid samples, or the like, as well as any qualitycontrol materials and calibration solutions used in analyzer equipmentfor measuring any of these fluids.

Further according to some embodiments of the method, the fluid sample isa gas, e.g. a medical gas, such as a physiological gas. Correspondingly,a sample analyzer for use in the method is advantageously adapted toanalyzing parameters of medical gas samples. Examples of particularlyuseful medical gas samples are selected from the group of respiratorgas, expiratory air, or the like, as well as any quality control andcalibration materials used in analyzer equipment for measuring any ofthese fluids. A medical sample analyzer may thus advantageously includea gas handling system comprising valves, conduits, and/orpumping/transfer means, for controlling gas flow, such as for fillingand emptying of the measurement chamber with the gas sample—preferablyin an automated manner. Preferably, a medical sample analyzer comprisesfluid handling means suited for both liquid and gas.

A second aspect of the invention relates to an optical sensor for thedetection of a contaminant, the optical sensor comprising a sensor layerwith a sensor surface forming an interface to a sample space, stimulusmeans, detection means, data storage means, and a signal processor,wherein the sensor layer comprises a luminophor adapted to emitluminescence radiation in response to an excitation stimulus applied tothe luminophor; wherein the stimulus means is arranged for providing anexcitation stimulus to the luminophor in the sensor layer; wherein thedetection means is arranged to detect luminescence radiation emitted bythe luminophor in response to the excitation stimulus; wherein the datastorage means comprises programmed instructions for:

-   -   receiving a time sequence of measurement values for detected        luminescence as signals from the optical sensor as an input;    -   determining an actual value of a first parameter and an actual        value of a second parameter, based on the time sequence, wherein        one of the first and second parameters is sensitive to a change        in refractive index across the interface between the sensor        layer and the sample space, and wherein the other one of the        first and second parameters is not sensitive to said change in        refractive index across the interface between the sensor layer        and the sample space;    -   developing an expected value for the second parameter based on        the actual value of the first parameter;    -   performing a comparison of the expected value for the second        parameter to the actual value of the second parameter; and    -   determining presence or absence of a contaminant based on the        comparison;

and wherein the signal processor is operable to execute said programmedinstructions so as to produce an output indicative of the presence orabsence of a contaminant.

Thereby, a particularly compact and robust detection of contaminants onthe sensor surface is achieved, wherein the contaminants aredistinguishable from a fluid sample by their difference in refractiveindex as discussed above with respect to the method for detecting acontaminant on a sensor surface facing towards a sample space. Furtheradvantages of the optical sensor for the detection of a contaminant andfurther advantageous embodiments are, in an analogue manner, alsoevident from the above discussion of the method of detectingcontaminants and of additional features of the embodiments of the methoddisclosed herein. For example, the optical sensor is advantageouslyadapted to measure one or more analytes in a fluid sample so as todetermine a corresponding parameter of the analyte, such as pO₂, pCO₂,pH, concentrations of electrolytes, concentrations of metabolic factors,or concentrations of enzymes, as mentioned above.

The optical sensor, and advantageous embodiments thereof with theadditional features as disclosed in the context of the method, isparticularly useful for use in a measurement chamber of a fluid sampleanalyzer, preferably a medical sample analyzer.

In a third aspect, a measurement chamber comprises an optical sensoraccording to any of the embodiments disclosed herein. The measurementchamber is preferably adapted for testing for, or measuring parametersof, one or more analytes in a fluid sample. The optical sensor isarranged such that the sensor surface faces into a sample space definedby the measurement chamber.

In a fourth aspect, a fluid sample analyzer is adapted for performing amethod of contaminant detection according to any of the embodimentsdisclosed herein, the fluid sample analyzer comprising a measurementchamber with inlet and outlet ports for feeding and discharging a fluidsample to the measurement chamber and an optical sensor according to anyof the embodiments disclosed herein, the optical sensor being arrangedsuch that the sensor surface faces into a sample space defined by themeasurement chamber.

Further according to a fifth aspect, a computer-implemented method ofdetecting a contaminant in a measurement chamber of a sample analyzercomprises an optical sensor with a sensor layer comprising a luminophor,wherein the sensor layer has a sensor surface forming an interface tothe measurement chamber, the method comprising the steps of:

-   -   receiving a time sequence of measurement values representing        luminescence intensities as a function of time as detected in        response to a stimulus applied to the luminophor;    -   based on the time sequence, determining an actual value of a        first parameter and an actual value of a second parameter,        wherein one of the first and second parameters is sensitive to a        change in refractive index across the interface between the        sensor layer and the sample space, and wherein the other one of        the first and second parameters is not sensitive to said change        in refractive index across the interface between the sensor        layer and the sample space;    -   developing an expected value for the second parameter based on        the actual value of the first parameter;    -   comparing the expected value for the second parameter to the        actual value of the second parameter; and    -   determining the presence or absence of a contaminant based on        the comparison.

Also in this aspect, further advantageous embodiments and any advantagesoriginating from these are in an analogue manner based on the additionalfeatures as discussed above with respect to the optical sensor and themethod of detecting a contaminant.

Further according to a sixth aspect, a software product that can beloaded to a processor, the processor being configured for communicatingwith an optical sensor comprising a sensor layer, the sensor layercomprising a sensor surface facing towards a sample space, the sensorlayer comprising a luminophor, the processor being further configuredfor controlling stimulus means adapted for exciting the luminophor, thesoftware product comprising instructions for:

-   -   (i) operating stimulus means to apply a stimulus to the        luminophor in the sensor layer;    -   (ii) operating the optical sensor to detect luminescence emitted        from the luminophor in the sensor layer in response to the        stimulus as a function of time;    -   (iii) obtaining a time sequence of measurement values for the        detected luminescence;    -   (iv) based on the time sequence, determining an actual value of        a first parameter and an actual value of a second parameter,        wherein one of the first and second parameters is sensitive to a        change in refractive index across the interface between the        sensor layer and the measurement chamber, and wherein the other        one of the first and second parameters is not sensitive to said        change in refractive index across the interface between the        sensor layer and the measurement chamber;    -   (v) developing an expected value for the second parameter based        on the actual value of the first parameter;    -   (vi) performing a comparison of the expected value for the        second parameter to the actual value of the second parameter;        and    -   (vii) determining the presence or absence of a contaminant on        the sensor surface based on the comparison.

Also in this aspect, further advantageous embodiments and any advantagesoriginating from these are in an analogue manner based on the additionalfeatures as discussed above with respect to the optical sensor and themethod of detecting a contaminant.

BRIEF DESCRIPTION OF THE DRAWINGS

Preferred embodiments of the invention will be described in more detailin connection with the appended drawings, which show in

FIG. 1 a diagram of a blood analyzer according to one embodiment;

FIG. 2 schematically, an optical sensor according to one embodiment;

FIG. 3 schematically, a detail of the optical sensor of FIG. 2; and in

FIG. 4 a graph with the time-dependence of the luminescence intensityfor two different sample fluids as detected with an optical sensoraccording to one embodiment.

FIG. 5 shows the principle of bubble and clot check.

FIG. 6a shows the result from pO2 measurement on liquid samples withoutand with an air bubble.

FIG. 6b shows the bubble factor for five samples of which four (sample1, 3, 4, and 5) do not have a bubble and one (sample 2) has a bubble.

FIG. 7a shows the result from pO2 measurement on gas samples without andwith a clot.

FIG. 7b shows the clot factor for five samples of which four (sample 1,2, 4, and 5) do not have a clot and one (sample 3) has a clot.

DETAILED DESCRIPTION

FIG. 1 shows schematically a liquid sample analyzer 1 with an analyzerpart having a signal processor 8, one or more analyte sensors 3(a-i), 4,a measurement chamber 2, and fluid handling infrastructure 20. Forperforming measurements, the user may provide a liquid sample at aninput port 12 a/b of the analyzer 1. The liquid sample is transferredthrough an inlet port 6 to the measurement chamber 2 comprising aplurality of analyte sensors 3, 4. The analyte sensors 3, 4 are arrangedto provide essentially simultaneous measurements on analyte parametersin a liquid sample, e.g. a whole blood sample. Preferably, the requiredsample amount for obtaining precise and reliable data is as small aspossible. A detailed example of a sensor assembly design that isparticularly suitable for simultaneously measuring a plurality ofdifferent parameters in bodily fluids, particularly in whole blood, andits use in a blood analyzer is e.g. found in EP 2 147 307 B1. Followingpre-programmed instructions loaded in a signal processor 8 and/or userinput, measurements are performed using the analyte sensors 3, 4. Theanalyte sensors 3, 4 generate signals that are representative of aphysical parameter for the respective analyte and provide the signals tothe signal processor 8 of the analyzer part. The signal processor 8 isadapted to receive and process signals from the analyte sensors 3, 4,and present the processed signals as output to a user or to asubsequent/further data analysis. After measurement, the liquid sampleis discharged, and the measurement chamber 2 is prepared for the nextmeasurement. The embodiment of the analyzer shown in FIG. 1 isparticularly adapted for the measurement of blood parameters, andfurther comprises an optional oxygenation measurement device 9downstream of the measurement chamber 2. Performing the measurements,calibration tasks, and quality control procedures thus typicallyinvolves the loading, unloading, rinsing, cleaning and re-loading ofdifferent liquids, which may be done by the fluid handlinginfrastructure 20. The fluid handling may be controlled in an automatedway by the signal processor 8 according to pre-programmed instructionsand/or user input. The fluid handling infrastructure 20 includes anumber of reservoirs 21 pre-filled with process liquids (RINSE/CAL1,CAL2, QC1, QC2, QC3) for rinsing/wash-out, calibration and qualitycontrol tasks. The process liquids (RINSE/CAL1, CAL2, QC1, QC2, QC3)have a known composition. The exact composition of a given batch may bestored in a chip 25 that may be attached to a cassette comprising thereservoirs 21, wherein the chip 25 may be read by the signal processor8. The process liquid (RINSE/CAL1, CAL2, QC1, QC2, QC3) for a givenprocess step may be selected by a fluid selector valve 22, and via feedline 12 c transferred through the inlet port 6 to the measurementchamber 2. Correct filling of the measurement chamber 2 may be monitoredand verified by visual inspection or according to known procedures byobserving the propagation of a liquid interface through the system bymeans of liquid sensors 10 a, 10 b, 10 c located upstream and downstreamof the measurement chamber, such as at the inlet 6 (“LS inlet” 10 a), atthe outlet 7 (“LS BG” 10 b), and just after the oxygenation measurementdevice 9 (“LS OXI” 10 c), respectively. The fluid flow through theanalyzer is driven by a pump 23, here a peristaltic hose-pump arrangeddownstream of the measurement chamber 2 and the oxygenation measurementdevice 9 and connected thereto via fluid line 13. The discharged fluidsare finally transported through fluid line 14 to the waste reservoir 24.

Upon start-up and, in an ongoing manner, during uptime, the analyzer 1performs self-control routines. If any abnormality is detected, theanalyzer 1 indicates the deviation to a user, and may further indicateways of overcoming an error state. On the other hand, when the analyzerindicates normal operation, measurements can be performed immediately.Advantageously according to some embodiments, the self-control routinesmay be performed during idle times, i.e. when the analyzer is in an idlestate, where it is not used for performing actual measurements on auser's sample. The self-control routines may include continuedrepetitive measurements performed on a calibration-grade process liquidwith a precisely known composition, as e.g. stored on chip 25. Thesignals obtained for each of the different analyte sensors 3, 4 on thewell-known composition may then be used to continuously update thereference for the respective analyte measurements.

Turning to FIG. 2 and FIG. 3, an embodiment of an optical detectorset-up is now discussed. FIG. 2 shows schematically an optical detector200 according to one embodiment in contact with a measurement chamber201 defined by sidewalls 202, 204. The optical detector 200 is adaptedfor measuring a physical parameter of an analyte in a fluid sample 99placed in the measurement chamber 201, such as a partial pressure of ananalyte-gas. The optical detector 200 has a sensor layer 205 with asensor surface 206 facing towards the inside of the measurement chamber201. The sensor surface 206 thus forms an interface to the measurementchamber 201 receiving the fluid sample 99, wherein the sensor surface206 is adapted for directly contacting the fluid sample 99. Thearrangement of the sensor layer 205 in the wall 202 of the measurement201 is best seen in FIG. 3, which shows a detail of the part of theoptical detector 200 that is integrated in the wall 202. In thisparticular embodiment, the wall 202 is made of a substrate 220, madee.g. of ceramics, carrying encapsulant and polymer layers 221, 222, 223.A thinned portion 224 of the substrate 220 allows for optical access tothe sensor layer 205. The sensor layer 205 is applied to the front sideof the substrate 220, i.e. to the side facing towards the measurementchamber 201, within an opening in the encapsulant and polymer layers221, 222, 223 that is also aligned with the thinned portion 224. In thisarrangement, the sensor layer 205 response can be probed optically fromthe backside of the substrate 220, i.e. from the side facing away fromthe measurement chamber 201, through the thinned portion 224 functioningas a window like optical access. Note that for reasons of clarity, thelayer thicknesses shown in FIG. 3 are not to scale. Suitable thicknessesfor a window portion 224 may be chosen, e.g. according to considerationsof mechanical strength and optical transparency. Suitable thicknessesfor a sensor layer 205 may be chosen, e.g. according to considerationsof response time and of establishing an equilibrium state of the sensorlayer 205 during measurements within a desired time frame, such aswithin 1 s, within 3 s, within 10 s, within 30 s, or within 1 min. Theseconsiderations may take into account, for example, the diffusivity ofthe relevant analyte into and out of the sensor layer 205. For example,the window portion 224 of the ceramic substrate 220 may have a thicknessof about 100 μm, whereas a typical layer thickness of the sensor layer205 may be in the range between 1 μm and 10 μm, typically between 1 μmand 4 μm, or about 2.5 μm.

The sensor layer 205 comprises a luminophor 210, i.e. a materialemitting luminescence I(t) in response to an excitation-stimulus S(t).The excitation stimulus S(t) is typically provided in the form of pulsedor modulated light generated by a suitable light source 207, such as alight source using a light emitting diode or a laser, in an excitationwavelength range adapted to optically excite the luminophor 210 into anexcited state from which it then decays under emission of radiation inan emission wave length range that is spectrally distinguishable fromthe excitation wavelength range. In the embodiment shown in FIG. 2,excitation light S(t) from an LED 207 is coupled by a mirror 208 atnormal incidence onto the back side of the substrate 220, thusilluminating the sensor layer 205 through the thinned portion 224. Theincident excitation light S(t) excites a fraction of the luminophormolecules 210 in the sensor layer 205, which responds by emittingluminescence light in essentially all directions, as indicated by thestar symbols in FIG. 3. Typically, the luminophor is of thedown-conversion type, i.e. the luminophor emission light is found atlower photon energies, or longer wavelengths, as compared to theexcitation light. However, up-converting luminophor substances, i.e.substances where the luminophor emission light is found at higher photonenergies, or shorter wavelengths, as compared to the excitation light,are also conceivable as long as the response radiation R(t) isspectrally distinguishable from the stimulus radiation S(t). Lightemitted from the luminophor 210 is collected from the back side of thesubstrate 220, through the thinned portion 224 as response radiationR(t). The response radiation R(t) is then separated from the stimulusradiation S(t) by adequate spectral separation means, such as by meansof a dichroic mirror 208. The response radiation R(t) is then detectedby a photodetector 209, which generates a sensor output signal that canbe amplified, acquired and processed in any suitable way by any suitableanalog and/or digital data processing means as known in the art ofsensor signal processing and data acquisition.

The intensity of the response radiation R(t) that is collected anddetected by the detector 209 depends on the optical properties of thespecific detection set-up, and in particular on the optical elements onthe optical path from the luminophor 210 in the sensor layer 205 to thephotodetector 209. Due to this dependence on factors extrinsic to theluminescence process, the collected and detected intensity of theresponse radiation R(t) may be called an “extrinsic” parameter. Forexample, the intensity of the collected response radiation R(t) dependson the difference in refractive index across the interface 206 betweenthe sensor layer 205 having refractive index n₁ and the fluid samplehaving refractive index n₂ as best seen in FIG. 3. A luminophor molecule210 that is excited by incident stimulus radiation S(t) emitsluminescence in all directions. Luminescence light travelling in adirection away from the substrate 220 towards the fluid sample 99 ispartly reflected at the interface 206 and partly transmitted into thefluid sample 99. The reflected part contributes to the collected anddetected intensity of the response radiation R(t), whereas thetransmitted part of the luminescence intensity is lost for detection.The effect of the interface 206 on the collection efficiency depends onthe difference in refractive index between the sensor layer 205 (n₁) andthe adjacent fluid sample 99 (n₂). As a consequence, the intensity ofthe collected and detected radiation R(t) is sensitive to changes in therefractive index n₂ of the fluid sample 99 in contact with the sensorsurface 206. Keeping remaining configuration parameters of the opticaldetector set-up 200 comparable to each other, the intensity of thecollected and detected radiation originating from the luminophor 210 inthe sensor layer 205 can thus be used to discriminate between differentsample substances that are distinguishable by their refractive indices,when these are in contact with the sensor surface 206. For example, theoptical detector set-up 200 may be used to distinguish between a fluidsample 99 of an aqueous solution having a refractive index of about 1.3and a fluid sample 99 of a gas having a refractive index of 1, whereinthe sensor layer 205 includes e.g. a polymer matrix material hosting theluminophor 210, the matrix material having e.g. a refractive index of1.5.

For a given optical detector set-up, the change in intensity to beexpected from a given change in the refractive index of the fluid sample99 may be determined empirically by routine experimentation. By way ofexample, for an optical detector setup 200 as described above, the ratioR(liquid)/R(gas) of the collected and detected intensities R_(gas)(t)and R_(liquid)(t) for a gas sample and for an aqueous solution presentedat the interface 206, respectively, is determined asR(liquid)/R(air)=0.52. Since the luminophor employed is typically alsosensitive to the presence of a specific analyte, here for example formeasuring the partial pressure of oxygen pO₂ in the fluid sample 99,care has been taken to determine the ratio R(liquid)/R(gas) forcomparable gas and liquid samples, i.e. gas and liquid samples with thesame concentration of the specific analyte. For example, in the case ofa gas analyte, such as oxygen, a gas sample with a specific partialpressure for that gas analyte may be selected as the gas sample, and acorresponding liquid sample of an aqueous solution with the same partialpressure of the gas analyte can be prepared by aerating the aqueoussolution with gas of the same composition (e.g. using gas from the samesource) as the gas sample until an equilibrium concentration isestablished. In so far the ratio of the intensities for R(liquid)/R(gas)is taken for comparable liquid and gas samples at a given point in time,the ratio is observed to be independent of the analyte concentration,which supports the extrinsic nature of the response radiation intensityas a parameter derivable from the collected and detected luminescenceresponse.

The stimulus radiation S(t) is typically provided in one or more pulses.Typically, a sequence of pulses is provided, wherein the pulses areseparated by idle periods where the stimulus is switched off. For alight source using LED emitters, the stimulus radiation is typicallymodulated in time as a sequence of ON-states separated by OFF-states,which may be considered as pulses and idle periods, respectively. Aftertermination of the stimulus S(t), e.g. when switching from an ON-stateto an OFF-state, the intensity of the luminescence I(t) decreases with atime constant that is characteristic for the luminophor, also referredto as the lifetime τ₀. The observed lifetime τ₀ is intrinsic to therecombination processes occurring in the luminophor. The lifetime τ₀, assuch, is therefore not affected by the particular configuration of theoptical path of the emitted light from the luminophor to the detector,such as the presence of interfaces between optical elements (e.g.layers), or the refractive index or optical density of such opticalelements (e.g. layers). The lifetime τ₀ may therefore be referred to asan “intrinsic” parameter, which is derivable from the collected anddetected luminescence response R(t).

For a given set-up for the optical detection, fluid samples with thesame analyte concentration will yield the same result for the lifetimeτ. If the samples also have the same refractive index, the sameintensity signal is expected at a given point in time after terminationof the stimulus radiation.

In order for the optical detector 200 to function as a sensor for aspecific analyte, the luminophor 210 is sensitive to the presence of thespecific analyte, wherein the luminescence R(t) as collected anddetected in response to the stimulus S(t) is a function of theconcentration of the analyte in the fluid sample 99 in contact with thesensor surface 206. The optical detector is operated to generate acorresponding output signal, which can be analyzed to provide a measurefor the concentration of the analyte. The underlying sensor principlemay be for example the quenching of the luminescence emission from theexcited luminophor by the presence of the analyte acting as a so-calledquenching agent. As a consequence of the quenching mechanism, thelifetime τ₀ and the intensity of the luminescence at a given point intime is a function of the analyte concentration.

Typically, the luminophors employed are phosphors with a lifetime τ₀above 1 μs in order to facilitate an easy detection of thetime-dependence of the emitted luminescence R(t) with relatively simpleinstrumentation. For example, the luminophor may be apalladiumporphyrin, e.g. palladium(II)-tetraphenylporphyrin (PdTPP) orpalladium(II)-tetra-(pentafluorophenyl)-porphyrin (PdTFPP), or any othersuitable luminophor. Palladiumporphyrins are, for example, well suitedfor blood measurements when immobilized in a polymer matrix to form asensor layer 205.

The analyte concentration of a fluid sample, such as the partialpressure of oxygen pO₂, may be determined from the observed decrease inlifetime due to luminescence quenching using the Stern-Volmer equation:

${{{pO}_{2}(\tau)} = {k*\left( {\frac{\tau_{0}}{\tau} - 1} \right)}},$

wherein τ₀ is the lifetime of the luminescence response R(t) for asample with zero concentration, e.g. an argon gas sample. Thesensitivity coefficient k may be determined from measurements on one ormore samples with known analyte concentrations, for example anatmospheric air sample with 21% of oxygen at normal conditions/correctedfor barometric pressure, or a calibration liquid (aqueous solution) thathas been prepared with a predefined partial pressure of O₂, e.g. in therange between 50-250 mmHg, or a corresponding gas sample prepared withthe same partial pressure of O₂.

FIG. 4 shows a graph with the time-dependence of the luminescenceintensity for two different fluid samples as detected with an opticalsensor according to one embodiment, namely for a gas sample and a liquidsample, respectively. The graph shows a plot of the luminescenceintensity on the coordinate axis vs. time on the ordinate axis. The plothas a first, excitation phase “I”, during which a stimulus radiation isin an ON-state, and a second, idle phase “II”, during which the stimulusradiation is in an OFF-state. The first phase starts just before theordinate value “−250”, when the stimulus radiation is switched on, andterminates at the ordinate value “0”, when the stimulus radiation isswitched off again. The stimulus radiation pulse of phase “I” causes theluminescence emission to increase as a larger and larger fraction of theluminophor in the sensor layer is excited by the incident stimulus andsubsequently relaxes through radiative processes. The increase inluminescence intensity tends to saturate as the excitation processes andthe relaxation processes compete with each other. The second phasestarts at the ordinate value “0”, upon termination of the first phase.The second phase extends as long as the stimulus remains in theOFF-state, typically until the next excitation pulse starts. Upontermination of the stimulus radiation pulse, the luminescence intensitydecreases as the fraction of excited luminophor decreases due torelaxation. The decrease is characterized by a lifetime τ, which dependson the concentration of analyte in the two fluid samples.

Since both fluid samples have been prepared with the same analyteconcentration and measurements, an analysis of the time dependence inthe second phase of the plot yields the same characteristic lifetime forboth samples. The difference in the observed intensity is a consequenceof the difference in refractive index between the two samples, which areabout 1.3 for the liquid sample and about 1 for the gas sample, whereinthe sensor layer has a refractive index of about 1.5.

The optical detector used in the example is for measuring partialpressure of oxygen pO₂ in the fluid samples using an analyzer asdescribed above with respect to FIG. 1. The gas sample of FIG. 4 is areference gas that has been prepared with an oxygen-content at a partialpressure of approximately 500 mmHg and the liquid sample of FIG. 4 is anaqueous solution that has been aerated to an equilibrium state with agas of the identical composition as the gas sample. The two measuredsample fluids, gas and liquid, are thus prepared with the same partialpressure of oxygen pO₂ contained therein. The two gas and liquid samplesare therefore at least comparable with respect to the analyte to whichthe optical detector is sensitive (here: oxygen).

FIG. 5 shows the principle of bubble and clot check by pO2 measurement.The solid line is the calibration on a known gas (i.e. a reference gas)from where Energy gas, E(g) and tau gas, T(g) are determined. The dottedline is measurement on a liquid sample from where Energy liquid, E(I)and tau liquid, T(I) are determined. Data presented in the graph are pO2measurement at the same pO2 level, i.e. with the same level of tau.

Bubble Check: BubbleFactor=Energy liquid/Tau liquid/(Energy gas/Tau gas)

-   -   Nominal value liquid=LiquidFactor

LiquidFactor=Energy liquid/Energy gas=0.52 is empirically determined onthe overall pO2 system and applies to all levels of pO2.

Clot Check: ClotFactor=Energy gas/Tau gas/(Energy gas/Tau gas)

-   -   Nominal value gas=1

The nominal value is the mean value which has been observed for a fluidsample without the presence of a bubble or clot. It is used forcalculating the threshold, i.e. the upper and lower acceptance limits.

EXAMPLE 1

The result from pO2 measurement on Liquid samples without and with anair bubble is shown in FIGS. 6a & 6 b. FIG. 6a shows the intensity overtime for a sample without (solid line) and with (dotted line) a bubble.FIG. 6b shows the bubble factor for five samples of which four (sample1, 3, 4, and 5) do not have a bubble and one (sample 2) has a bubble.

TABLE 1 Calibration of the sensor and calculation of bubble factor forsample 2 & 3. Calibration of the sensor on gas with a level of 140 mmHgE (g) 1024 count T (g) 2.50E−04 microsec Calculation of bubble factor ona sample with a bubble with a level of 70 mmHg (sample 2) E (l) 1300count T (l) 4.00E−04 microsec Calculation of bubble factor on a samplewithout bubble with a level of 70 mmHg (sample 3) E (l)  850 count T (l)4.00E−04 microsec

EXAMPLE 2

The result from pO2 measurement on gas samples without and with a clotis shown in FIGS. 7a & 7 b. FIG. 7a shows the intensity over time for asample without (solid line) and with (dotted line) a clot. FIG. 7b showsthe clot factor for five samples of which four (sample 1, 2, 4, and 5)do not have a clot and one (sample 3) has a clot.

TABLE 2 Calibration of the sensor and calculation of clot factor forsample 2 & 3. Calibration of the sensor on gas with a level of 140 mmHgE (g) 1024 count T (g) 2.50E−04 microsec Calculation of clot factor fora sample without a clot with a level of 175 mmHg (sample 2) E (g)  805count T (g) 2.10E−04 microsec Calculation of clot factor for a samplewith a clot with a level of 175 mmHg (sample 3) E (g)  610 count T (g)2.10E−04 microsec

1-27. (canceled)
 28. A method of detecting a contaminant in ameasurement chamber of a sample analyzer, wherein the sample analyzercomprises an optical sensor with a sensor layer comprising a luminophor,wherein the sensor layer has a sensor surface forming an interface to afluid sample present in the measurement chamber, the method comprising:a. filling the measurement chamber with a fluid sample; b. applying astimulus to the luminophor in the sensor layer; c. detectingluminescence emitted from the luminophor in the sensor layer in responseto the stimulus as a function of time; d. obtaining a time sequence ofmeasurement values for the detected luminescence; e. based on the timesequence, determining an actual value of a first parameter and an actualvalue of a second parameter, wherein one of the first and secondparameters is sensitive to a change in refractive index across theinterface between the sensor layer and the measurement chamber, andwherein the other one of the first and second parameters is notsensitive to said change in refractive index across the interfacebetween the sensor layer and the measurement chamber; and f. providingfeedback on a contamination state based on the actual value of the firstparameter and the actual value of the second parameter.
 29. The methodaccording to claim 28, wherein providing feedback on a contaminationstate based on the actual value of the first parameter and the actualvalue of the second parameter comprises: g. developing an expected valuefor the second parameter based on the actual value of the firstparameter; h. comparing the expected value for the second parameter tothe actual value of the second parameter; and i. determining thepresence of a contaminant based on the comparison or determining theabsence of a contaminant based on the comparison.
 30. The methodaccording to claim 28, wherein providing feedback on a contaminationstate based on the actual value of the first parameter and the actualvalue of the second parameter comprises: g. developing an actual valueof a third parameter based on the actual value of the first parameterand the actual value of the second parameter; h. comparing the actualvalue of the third parameter with an expected value for the thirdparameter; and i. determining the presence of a contaminant based on thecomparison or determining the absence of a contaminant based on thecomparison.
 31. The method according to claim 29, wherein presence of acontaminant is determined if the difference between the actual value ofthe second parameter and the expected value for the second parameter isabove a threshold, and/or wherein absence of a contaminant is determinedif the difference between the actual value of the second parameter andthe expected value for the second parameter is below the threshold. 32.The method according to claim 30, wherein presence of a contaminant isdetermined if the difference between the actual value of the thirdparameter and the expected value for the third parameter is above athreshold, and/or wherein absence of a contaminant is determined if thedifference between the actual value of the third parameter and theexpected value for the third parameter is below the threshold.
 33. Themethod according to claim 28, further comprising providing referenceinformation, and wherein providing feedback on a contamination statebased on the actual value of the first parameter and the actual value ofthe second parameter is further based on the reference information. 34.The method according to claim 33, wherein providing referenceinformation comprises performing reference measurements for one or moreknown contamination states.
 35. The method according to claim 28,wherein the fluid sample is an aqueous liquid or a gas.
 36. The methodaccording to claim 28, wherein the fluid sample is a liquid with arefractive index between 1.2 and 1.5, and/or wherein the refractiveindex of the sensor layer is at least 1.4.
 37. The method according toclaim 28, wherein the obtaining a time sequence of measurement valuesincludes measuring the luminescence intensity at a plurality of at leastthree points in time.
 38. The method according to claim 28, wherein thefirst parameter corresponds to the lifetime τ of the luminescence,and/or wherein the second parameter corresponds to the intensity of theluminescence at a given point in time.
 39. The method according to claim28, wherein the luminophor in the sensor layer is a phosphor with aluminescence lifetime between 1 μs and 1 s; and/or wherein theluminophor in the sensor layer is a phosphor with a luminescencelifetime of at least 10 μs; and/or wherein the luminophor in the sensorlayer is a phosphor with a luminescence lifetime up to and including 1s.
 40. The method according to claim 28, wherein the filling themeasurement chamber includes bringing the fluid sample in a diffusiveequilibrium with the sensor layer, at least with respect to one analyte.41. The method according to claim 28, wherein the optical sensor isadapted for measuring a partial pressure of a gas fraction in the fluidsample.
 42. The method according to according to claim 28, wherein thesensor layer is adapted for the diffusive uptake of an analyte from thefluid sample, and wherein the luminophor in the sensor layer issusceptible to luminescence quenching due to the presence of the analytein the sensor layer.
 43. The method according to claim 28, wherein theoptical sensor is adapted to determine a parameter of one or moreanalytes, wherein the parameter of the one or more analytes is selectedfrom the group of: pO2, pCO2, pH; concentrations of electrolytes;concentrations of metabolic factors; and concentrations of enzymes. 44.The method according to claim 28, wherein the fluid sample is a liquidselected from the group of blood, diluted or undiluted whole blood,serum, plasma, saliva, urine, cerebrospinal liquid, pleura, synovialliquid, ascites liquid, peritoneal liquid, amniotic liquid, milk, anddialysis liquid samples, or wherein the fluid sample is a medical gassample selected from the group of respirator gas or expiratory air. 45.An optical sensor for the detection of a contaminant, the optical sensorcomprising a sensor layer with a sensor surface forming an interface toa sample space, stimulus means, detection means, data storage means, anda signal processor, wherein the sensor layer comprises a luminophoradapted to emit luminescence radiation in response to an excitationstimulus applied to the luminophor; wherein the stimulus means isarranged for providing an excitation stimulus to the luminophor in thesensor layer; wherein the detection means is arranged to detectluminescence radiation emitted by the luminophor in response to theexcitation stimulus; wherein the data storage means comprises programmedinstructions for: receiving a time sequence of measurement values fordetected luminescence as signals from the optical sensor as an input;determining an actual value of a first parameter and an actual value ofa second parameter, based on the time sequence, wherein one of the firstand second parameters is sensitive to a change in refractive indexacross the interface between the sensor layer and the sample space, andwherein the other one of the first and second parameters is notsensitive to said change in refractive index across the interfacebetween the sensor layer and the sample space; providing feedback on acontamination state based on the actual value of the first parameter andthe actual value of the second parameter, wherein the signal processoris operable to execute said programmed instructions so as to produce anoutput indicative of the feedback on a contamination state.
 46. A fluidsample analyzer adapted for performing a method of contaminant detectionaccording to claim 28, the fluid sample analyzer comprising ameasurement chamber with inlet and outlet ports for feeding anddischarging a fluid sample to the measurement chamber and an opticalsensor for the detection of a contaminant, the optical sensor comprisinga sensor layer with a sensor surface forming an interface to a samplespace, stimulus means, detection means, data storage means, and a signalprocessor, wherein the sensor layer comprises a luminophor adapted toemit luminescence radiation in response to an excitation stimulusapplied to the luminophor; wherein the stimulus means is arranged forproviding an excitation stimulus to the luminophor in the sensor layer;wherein the detection means is arranged to detect luminescence radiationemitted by the luminophor in response to the excitation stimulus;wherein the data storage means comprises programmed instructions for:receiving a time sequence of measurement values for detectedluminescence as signals from the optical sensor as an input; determiningan actual value of a first parameter and an actual value of a secondparameter, based on the time sequence, wherein one of the first andsecond parameters is sensitive to a change in refractive index acrossthe interface between the sensor layer and the sample space, and whereinthe other one of the first and second parameters is not sensitive tosaid change in refractive index across the interface between the sensorlayer and the sample space; providing feedback on a contamination statebased on the actual value of the first parameter and the actual value ofthe second parameter, wherein the signal processor is operable toexecute said programmed instructions so as to produce an outputindicative of the feedback on a contamination state, and wherein theoptical sensor is arranged such that the sensor surface faces into asample space defined by the measurement chamber.
 47. A software productthat can be loaded to a processor, the processor being configured forcommunicating with an optical sensor comprising a sensor layer, thesensor layer comprising a sensor surface facing towards a sample space,the sensor layer comprising a luminophor, the processor being furtherconfigured for controlling stimulus means adapted for exciting theluminophor, the software product comprising instructions for: (i)operating stimulus means to apply a stimulus to the luminophor in thesensor layer; (ii) operating the optical sensor to detect luminescenceemitted from the luminophor in the sensor layer in response to thestimulus as a function of time; (iii) obtaining a time sequence ofmeasurement values for the detected luminescence; (iv) based on the timesequence, determining an actual value of a first parameter and an actualvalue of a second parameter, wherein one of the first and secondparameters is sensitive to a change in refractive index across theinterface between the sensor layer and the measurement chamber, andwherein the other one of the first and second parameters is notsensitive to said change in refractive index across the interfacebetween the sensor layer and the measurement chamber; (v) providingfeedback on a contamination state based on the actual value of the firstparameter and the actual value of the second parameter.